IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ PI 3-kinase: structural and functional analysis of intersubunit interactions. PI 3-kinase: structural and functional analysis of intersubunit interactions. EMBO J. (0261-4189) 1994 imex curation Dhand R., Hara K., Hiles I., Bax B., Gout I., Panayotou G., Fry MJ., Yonezawa K., Kasuga M., Waterfield MD. No contact email provided Lysates of Sf9 cells coinfected with p85alpha or beta and p110 viruses were precipitated with antibodies to p85alpha or beta. PI 3-kinase assays were performed on immunoprecipitates using phosphatidylinositol as a substrate. The reference Yonezawa et al. (1992) confirmed that p85beta was of bovine origin. The reference Hiles et al. (1992) gives sequence information that confirms that it is the alpha isoform of p110 that was used. DAVET Only protein-protein interactions in vitro In vitro enzymatic study enzymatic study weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Dhand R., Hara K., Hiles I., Bax B., Gout I., Panayotou G., Fry MJ., Yonezawa K., Kasuga M., Waterfield MD. No contact email provided Lysates of Sf9 cells coinfected with p85alpha or beta and p110 viruses were precipitated with antibodies to p85alpha or beta. PI 3-kinase assays were performed on immunoprecipitates using phosphatidylinositol as a substrate. The reference Yonezawa et al. (1992) confirmed that p85beta was of bovine origin. The reference Hiles et al. (1992) gives sequence information that confirms that it is the alpha isoform of p110 that was used. Accepted 2007-DEC-19 by DAVET. EMBO J. (0261-4189) 1994 DAVET Only protein-protein interactions imex curation PI 3-kinase: structural and functional analysis of intersubunit interactions. PI 3-kinase: structural and functional analysis of intersubunit interactions. EMBO J. (0261-4189) 1994 imex curation Dhand R., Hara K., Hiles I., Bax B., Gout I., Panayotou G., Fry MJ., Yonezawa K., Kasuga M., Waterfield MD. No contact email provided Lysates of Sf9 cells coinfected with p85alpha or beta and p110 viruses were precipitated with antibodies to p85alpha or beta. PI 3-kinase assays were performed on immunoprecipitates using phosphatidylinositol as a substrate. The reference Yonezawa et al. (1992) confirmed that p85beta was of bovine origin. The reference Hiles et al. (1992) gives sequence information that confirms that it is the alpha isoform of p110 that was used. DAVET Only protein-protein interactions spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection anti bait coip anti bait coimmunoprecipitation anti bait coip weight by comassie molecular weight estimation by coomasie staining weight by comassie Dhand R., Hara K., Hiles I., Bax B., Gout I., Panayotou G., Fry MJ., Yonezawa K., Kasuga M., Waterfield MD. No contact email provided Lysates of Sf9 cells coinfected with p85alpha or beta and p110 viruses were precipitated with antibodies to p85alpha or beta. PI 3-kinase assays were performed on immunoprecipitates using phosphatidylinositol as a substrate. The reference Yonezawa et al. (1992) confirmed that p85beta was of bovine origin. The reference Hiles et al. (1992) gives sequence information that confirms that it is the alpha isoform of p110 that was used. Accepted 2007-DEC-19 by DAVET. EMBO J. (0261-4189) 1994 DAVET Only protein-protein interactions imex curation PI 3-kinase: structural and functional analysis of intersubunit interactions. PI 3-kinase: structural and functional analysis of intersubunit interactions. EMBO J. (0261-4189) 1994 imex curation Dhand R., Hara K., Hiles I., Bax B., Gout I., Panayotou G., Fry MJ., Yonezawa K., Kasuga M., Waterfield MD. No contact email provided Lysates of Sf9 cells coinfected with p85alpha or beta and p110 viruses were precipitated with antibodies to p85alpha or beta. PI 3-kinase assays were performed on immunoprecipitates using phosphatidylinositol as a substrate. The reference Yonezawa et al. (1992) confirmed that p85beta was of bovine origin. The reference Hiles et al. (1992) gives sequence information that confirms that it is the alpha isoform of p110 that was used. DAVET Only protein-protein interactions in vitro In vitro enzymatic study enzymatic study weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Dhand R., Hara K., Hiles I., Bax B., Gout I., Panayotou G., Fry MJ., Yonezawa K., Kasuga M., Waterfield MD. No contact email provided Lysates of Sf9 cells coinfected with p85alpha or beta and p110 viruses were precipitated with antibodies to p85alpha or beta. PI 3-kinase assays were performed on immunoprecipitates using phosphatidylinositol as a substrate. The reference Yonezawa et al. (1992) confirmed that p85beta was of bovine origin. The reference Hiles et al. (1992) gives sequence information that confirms that it is the alpha isoform of p110 that was used. Accepted 2007-DEC-19 by DAVET. EMBO J. (0261-4189) 1994 DAVET Only protein-protein interactions imex curation PI 3-kinase: structural and functional analysis of intersubunit interactions. PI 3-kinase: structural and functional analysis of intersubunit interactions. EMBO J. (0261-4189) 1994 imex curation Dhand R., Hara K., Hiles I., Bax B., Gout I., Panayotou G., Fry MJ., Yonezawa K., Kasuga M., Waterfield MD. No contact email provided Lysates of Sf9 cells coinfected with p85alpha or beta and p110 viruses were precipitated with antibodies to p85alpha or beta. PI 3-kinase assays were performed on immunoprecipitates using phosphatidylinositol as a substrate. The reference Yonezawa et al. (1992) confirmed that p85beta was of bovine origin. The reference Hiles et al. (1992) gives sequence information that confirms that it is the alpha isoform of p110 that was used. DAVET Only protein-protein interactions in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Dhand R., Hara K., Hiles I., Bax B., Gout I., Panayotou G., Fry MJ., Yonezawa K., Kasuga M., Waterfield MD. No contact email provided Lysates of Sf9 cells coinfected with p85alpha or beta and p110 viruses were precipitated with antibodies to p85alpha or beta. PI 3-kinase assays were performed on immunoprecipitates using phosphatidylinositol as a substrate. The reference Yonezawa et al. (1992) confirmed that p85beta was of bovine origin. The reference Hiles et al. (1992) gives sequence information that confirms that it is the alpha isoform of p110 that was used. Accepted 2007-DEC-19 by DAVET. EMBO J. (0261-4189) 1994 DAVET Only protein-protein interactions imex curation p85b_bovin Phosphatidylinositol 3-kinase regulatory subunit beta Phosphatidylinositol 3-kinase 85 kDa regulatory subunit beta PIK3R2 protein protein bovin Bos taurus (Bovine) Bovine MAGPEGFQYRALYPFRRERPEDLELLPGDVLVVSRAALQALGVAEGNERCPQSVGWMPGLNERTRQRGDFPGTYVEFLGPVALARPGPRPRGPRPLPARPRDGPPEPGLTLPDLPEQFSPPDVAPPILVKLVEAIERTGLDSYRPEPPAVRTDWSLSDVEQWDAAALSDGVKGFLLALPAPLVTPEAAAEAHRALREAAGPVGPALEPPTLPLHHALTLRFLLQHLGRVAGRAPAPGPAVRALGATFGPLLLRAPPPPSPPPGGAPDGTEPTPDFPALLVEKLLQEHLEEQEVAPPALPPKPPKTKPAPTGLANGGSPPSLQDAEWYWGDISREEVNEKLRDTPDGTFLVRDASSKIQGEYTLTLRKGGNNKLIKVFHRDGHYGFSEPLTFCSVVDLITHYRHESLAQYNAKLDTRLLYPVSKYQQDQIVKEDSVEAVGAQLKVYHQQYQDKSREYDQLYEEYTRTSQELQMKRTAIEAFNETIKIFEEQGQTQEKCSKEYLERFRREGNEKEMQRILLNSERLKSRIAEIHESRTKLEQELRAQASDNREIDKRMNSLKPDLMQLRKIRDQYLVWLTQKGARQKKINEWLGIKNETEDQYSLMEDEDDLPHHEERTWYVGKINRTQAEEMLSGKRDGTFLIRESSQRGCYACSVVVDGDTKHCVIYRTATGFGFAEPYNLYGSLKELVLHYQHASLVQHNDALTVTLAHPVRAPGPGPPPAAR DIP protein P23726 original sequence version: 125 9D2BA8B6DB087098 1-phosphatidyl-1d-myo-inositol 1-phosphatidyl-1D-myo-inositols small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) pk3ca_bovin Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha isoform Phosphoinositide-3-kinase catalytic alpha polypeptide Serine/threonine protein kinase PIK3CA PIK3CA Phosphatidylinositol 4,5-bisphosphate 3-kinase 110 kDa catalytic subunit alpha protein protein bovin Bos taurus (Bovine) Bovine MPPRPSSGELWGIHLMPPRILVECLLPNGMIVTLECLREATLITIKHELFKEARKYPLHQLLQDESSYIFVSVTQEAEREEFFDETRRLCDLRLFQPFLKVIEPVGNREEKILNREIGFAIGMPVCEFDMVKDPEVQDFRRNILNVCKEAVDLRDLNSPHSRAMYVYPPNVESSPELPKHIYNKLDKGQIIVVIWVIVSPNNDKQKYTLKINHDCVPEQVIAEAIRKKTRSMLLSSEQLKLCVLEYQGKYILKVCGCDEYFLEKYPLSQYKYIRSCIMLGRMPNLMLMAKESLYSQLPMDCFTMPSYSRRISTATPYMNGETSTKSLWVINSALRIKILCATYVNVNIRDIDKIYVRTGIYHGGEPLCDNVNTQRVPCSNPRWNEWLNYDIYIPDLPRAARLCLSICSVKGRKGAKEEHCPLAWGNINLFDYTDTLVSGKMALNLWPVPHGLEDLLNPIGVTGSNPNKETPCLELEFDWFSSVVKFPDMSVIEEHANWSVSREAGFSYSHAGLSNRLARDNELRENDKEQLRAICTRDPLSEITEQEKDFLWSHRHYCVTIPEILPKLLLSVKWNSRDEVAQMYCLVKDWPPIKPEQAMELLDCNYPDPMVRGFAVRCLEKYLTDDKLSQYLIQLVQVLKYEQYLDNLLVRFLLKKALTNQRIGHFFFWHLKSEMHNKTVSQRFGLLLESYCRACGMYLKHLNRQVEAMEKLINLTDILKQEKKDETQKVQMKFLVEQMRRPDFMDALQGFLSPLNPAHQLGNLRLEECRIMSSAKRPLWLNWENPDIMSELLFQNNEIIFKNGDDLRQDMLTLQIIRIMENIWQNQGLDLRMLPYGCLSIGDCVGLIEVVRNSHTIMQIQCKGGLKGALQFNSHTLHQWLKDKNKGEIYDAAIDLFTRSCAGYCVATFILGIGDRHNSNIMVKDDGQLFHIDFGHFLDHKKKKFGYKRERVPFVLTQDFLIVISKGAQECTKTREFERFQEMCYKAYLAIRQHANLFINLFSMMLGSGMPELQSFDDIAYIRKTLALDKTEQEALEYFMKQMNDAHHGGWTTKMDWIFHTIKQHALN DIP protein P32871 original sequence version: 93 C753DCC2F39FDDF0 p85a_bovin Phosphatidylinositol 3-kinase regulatory subunit alpha Phosphatidylinositol 3-kinase 85 kDa regulatory subunit alpha PIK3R1 protein protein bovin Bos taurus (Bovine) Bovine MSAEGYQYRALYDYKKEREEDIDLHLGDILTVNKGSLVALGFSDGQEAKPEEIGWLNGYNETTGERGDFPGTYVEYIGRKKISPPTPKPRPPRPLPVAPGPSKTEADSEQQASTLPDLAEQFAPPDVAPPLLIKLVEAIEKKGLECSTLYRTQSSSNPAELRQLLDCDTASLDLEMFDVHVLADAFKRYLLDLPNPVIPVAVSSELISLAPEVQSSEEYIQLLKKLIRSPSIPHQYWLTLQYLLKHFFKLSQTSSKNLLNARVLSELFSPLLFRFPAASSENTEHLIKIIEILISTEWNERQPAPALPPKPPKPTTVANNGMNNNMSLQDAEWYWGDISREEVNEKLRDTADGTFLVRDASTKMHGDYTLTLRKGGNNKLIKIFHRDGKYGFSDPLTFNSVVELINHYRNESLAQYNPKLDVKLLYPVSKYQQDQVVKEDNIEAVGKKLHEYNTQFQEKSREYDRLYEDYTRTSQEIQMKRTAIEAFNETIKIFEEQCQTQERYSKEYIEKFKREGNETEIQRIMHNYEKLKSRISEIVDSRRRLEEDLKKQAAEYREIDKRMNSIKPDLIQLRKTRDQYLMWLTQKGVRQKKLNEWLGNENTEDQYSLVEDDEDLPHHDEKTWNVGSSNRNKAENLLRGKRDGTFLVRESSKQGCYACSVVVDGEVKHCVINKTATGYGFAEPYNLYSSLKELVLHYQHTSLVQHNDSLNVTLAYPVYAQQRR DIP protein P23727 original sequence version: 108 EBDF6E754BBF7321 pik3r1-pik3ca-1 2 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association 1 Ai In double infections of baculoviruses expressing p85a and p110, both proteins were detected in either anti-p85a or anti-p110 immunoprecipitates P23727; P32871 infection pik3ca-pik3r1-6 2 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association 1 Ai In double infections of baculoviruses expressing p85a and p110, both proteins were detected in either anti-p85a or anti-p110 immunoprecipitates P23727; P32871 infection pik3r2-pik3ca 2 7 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait physical association physical association 1 Ai P23726; P32871 infection Infected cells immunoprecipitated with antibodies directed to the p85 or the p1lO subunit, or if bound to Y751 phosphopeptide affinity beads, were found to contain both p850 and p110 proteins when analysed by SDS-PAGE pik3ca-pik3r2 2 5 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association 1 Ai P23726; P32871 infection Infected cells immunoprecipitated with antibodies directed to the p85 or the p1lO subunit, or if bound to Y751 phosphopeptide affinity beads, were found to contain both p850 and p110 proteins when analysed by SDS-PAGE pik3ca-1_phosphat-5 1 7 enzyme enzyme unspecified role unspecified role spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 6 enzyme target enzyme target substrate substrate unspecified role unspecified role 8 unspecified role unspecified role ancillary ancillary spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation 1 Aii pik3ca-1_phosphat-4 1 7 enzyme enzyme unspecified role unspecified role spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 5 unspecified role unspecified role ancillary ancillary spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 6 enzyme target enzyme target substrate substrate unspecified role unspecified role phosphorylation phosphorylation reaction phosphorylation 1 Aii pik3r1-pik3ca-2 4 7 unspecified role unspecified role prey prey spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 8 unspecified role unspecified role bait bait spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection physical association physical association 1 Bi Both p85a and p85,B were seen to bind p1 10 by Coomassie blue staining following SDS -PAGE pik3r2-pik3ca-1 4 5 unspecified role unspecified role bait bait spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 7 unspecified role unspecified role prey prey spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection physical association physical association 1 Bi Both p85a and p85,B were seen to bind p110 by Coomassie blue staining following SDS-PAGE pik3ca-pik3r1-7 4 7 unspecified role unspecified role bait bait spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 8 unspecified role unspecified role prey prey spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection physical association physical association 1 Biii Similar results were obtained when p110 was immobilized and the bound p85a protein was detected as a Coomassie blue-stained protein on SDS-PAGE following binding in vitro pik3ca-1_phosphat-6 3 5 unspecified role unspecified role ancillary ancillary spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 7 enzyme enzyme unspecified role unspecified role spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 6 enzyme target enzyme target substrate substrate unspecified role unspecified role phosphorylation phosphorylation reaction phosphorylation 1 Bii Both p85a and p85,B were seen to bind p110 by Coomassie blue staining following SDS-PAGE (Figure lBi, lanes 1 and 2) or upon assay for associated PI 3-kinase activity pik3ca-1_phosphat-7 3 8 unspecified role unspecified role ancillary ancillary spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 7 enzyme enzyme unspecified role unspecified role spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 6 enzyme target enzyme target substrate substrate unspecified role unspecified role phosphorylation phosphorylation reaction phosphorylation 1 Bii Both p85a and p85,B were seen to bind p110 by Coomassie blue staining following SDS-PAGE (Figure lBi, lanes 1 and 2) or upon assay for associated PI 3-kinase activity